Extended Data Fig. 7: In vitro assessment of self-renewal and differentiation potential in preleukemic cells from TP53-sAML patients.

a, Donor, type of clonal evolution and genotype of the 880 preLSCs identified. b, Proportion of HSCs in mobilized PB or BM from healthy donors (n = 7) and TP53-sAML patients (n = 9) in which preLSCs were detected. Graph shows mean ± s.e.m, “p” indicates two-tailed Student t-test p-value and “fc”, fold-change. c, Schematic representation of Lin⁻CD34⁺ cell fractions isolated and in vitro assays performed. TP53-sAML patient samples used (n = 3: IF0131, IF0391, GR001) were known to have TP53-WT preleukemic stem cells (preLSC) in the HSC compartment (Related to Fig. 3c). d-e, Long term culture-initiating cell in vitro assay. Percentage of positive wells in each immunophenotypic population (d) and clonogenic output (e) from healthy donor (HD, n = 4), MF (n = 3) and preLSCs from TP53-sAML (n = 3). Barplot indicates mean ± s.e.m. from 2 independent experiments. f, Representative cytospin images of HSC-derived colonies from the same patient groups as in (d-e). g, Genotyping of HSC and LMPP-derived colonies from the same LTC-IC assay as in (d-f), demonstrating absence of TP53 mutations in HSC-derived colonies, contrary to LMPPs. h, Percentage of CD34⁺ cells from healthy donor (HD, n = 4), MF (n = 3) and preLSCs from TP53-sAML (n = 2) after 12 days of liquid culture in conditions promoting hematopoietic differentiation. Barplot indicates mean ± s.e.m from 3 independent experiments, and “p”, two-tailed Student t-test p-value. i, Representative image of liquid culture HSC-derived colonies for healthy donor and TP53-sAML preLSCs, from the same experiment as in (h).